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The regulatory role of IL-6R in hepatitis B-associated fibrosis and cirrhosis - PubMed

  • ️Sun Jan 01 2017

The regulatory role of IL-6R in hepatitis B-associated fibrosis and cirrhosis

Y Chen et al. Braz J Med Biol Res. 2017.

Abstract

This study investigated the expression and regulation of IL-6R in hepatitis B-associated moderate hepatic fibrosis and cirrhosis. Liver tissues, peripheral blood monocytes (PBMs) and serum were collected from 26 hepatitis B patients with liver fibrosis and 35 hepatitis B patients with liver cirrhosis. The levels of Il-6r mRNA expression in these samples were examined by quantitative real-time PCR and IL-6R protein levels were analyzed by western blot and ELISA. MiRNAs that regulate IL-6R expression were predicted by bioinformatics analysis, and validated by dual luciferase reporter assay. Compared with the hepatic fibrosis group, IL-6R was significantly upregulated at both mRNA and protein levels in liver tissues, PBMs and serum samples from the hepatic cirrhosis group (P<0.05). The 3'UTR of Il-6r mRNA was predicted to contain a miR-30b binding site and IL-6R was identified as a possible target of miR-30b. MiR-30b expression was significantly downregulated in samples from hepatic cirrhosis patients compared with hepatic fibrosis patients (P<0.05). In conclusion, IL-6R was upregulated while miR-30b was decreased in patients with liver cirrhosis. The miR-30 can directly regulate the expression of IL-6R.

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Figures

Figure 1.
Figure 1.. Real-time PCR analysis of Il-6r mRNA expression. A, Liver tissues; B, peripheral blood monocytes (PBMs), and C, serum from hepatitis B patients with liver fibrosis or liver. Data are reported as means±SD. *P<0.05 and **P<0.01, one-way ANOVA.
Figure 2.
Figure 2.. Relative protein expression by western blot analysis of IL-6R protein expression. A, Liver tissues, and B, peripheral blood monocytes (PBMs) from hepatitis B patients with liver fibrosis (HP) or liver cirrhosis (HC). β-actin expression was detected as internal control. Data are reported as means±SD. *P<0.05 and **P<0.01, one-way ANOVA.
Figure 3.
Figure 3.. ELISA analysis of IL-6R protein expression in serum from hepatitis B patients with liver fibrosis and liver cirrhosis. Data are reported as means±SD. *P<0.05, ANOVA.
Figure 4.
Figure 4.. IL-6R is a possible target of miR-30b. The wildtype (WT) and mutant Il-6r 3′UTR luciferase reporter constructs were co-transfected with negative control RNA (NC) or agomiR-30b, respectively. Luciferase activities were assayed 24 h post transfection. Data are reported as means±SD. **P<0.01, compared with NC (one-way ANOVA).
Figure 5.
Figure 5.. Real-time PCR analysis of miR-30b expression in A, liver tissues, B, peripheral blood monocytes (PBMs), and C, serum from hepatitis B patients with liver fibrosis or liver cirrhosis. Data are reported as means±SD. *P<0.05 and **P<0.01, one-way ANOVA.

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